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[通过差异显示鉴定前庭代偿相关分子]

[Identification of vestibular compensation-associated molecules by means of differential display].

作者信息

Kitahara T, Takeda N, Okumura S, Kubo T

机构信息

Department of Otolaryngology, Osaka University Medical School.

出版信息

Nihon Jibiinkoka Gakkai Kaiho. 1998 Jan;101(1):37-43. doi: 10.3950/jibiinkoka.101.37.

Abstract

The differential display method was used to identify gene expression which is altered in the cerebellar flocculus after unilateral labyrinthectomy (UL). Total RNA from flocculi of sham-operated and labyrinthectomized rats was isolated, amplified by PCR using arbitrary primer sets and separated by electrophoresis on a polyacrylamide gel. PCR products, whose amounts were significantly different in samples from labyrinthectomized animals and those from controls, were cut out of the gel and sequenced. One of the up-regulated products was the rat protein phosphatase 2A beta catalytic subunit mRNA and one of the down-regulated products was the rat glutamate receptor delta-2 subunit mRNA. Histochemical examination of in situ hybridization showed that those molecules were intensively localized in the Purkinje cell layer. In labyrinthectomized rats, UL-induced nystagmus gradually disappeared within 3 days after UL. These findings suggest that changes in expression of those molecules in the floccular Purkinje cells after UL is involved in vestibular compensation. So far various kinds of neural plasticity-associated molecules have been investigated, mainly by slice-in vitro studies. This study indicates that differential display is a feasible molecular biological in vivo method for investigation of the mechanism of neural plasticity.

摘要

采用差异显示法来鉴定单侧迷路切除(UL)后小脑绒球中发生改变的基因表达。从假手术组和迷路切除大鼠的绒球中分离总RNA,使用任意引物组通过PCR进行扩增,并在聚丙烯酰胺凝胶上进行电泳分离。将在迷路切除动物样品和对照样品中含量有显著差异的PCR产物从凝胶中切下并测序。其中一个上调产物是大鼠蛋白磷酸酶2Aβ催化亚基mRNA,一个下调产物是大鼠谷氨酸受体δ-2亚基mRNA。原位杂交的组织化学检查表明,这些分子密集地定位于浦肯野细胞层。在迷路切除的大鼠中,UL诱导的眼球震颤在UL后3天内逐渐消失。这些发现表明,UL后绒球浦肯野细胞中这些分子表达的变化与前庭代偿有关。迄今为止,主要通过切片体外研究对各种神经可塑性相关分子进行了研究。本研究表明,差异显示是一种用于研究神经可塑性机制的可行的体内分子生物学方法。

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